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Modular glycosphere assays for high-throughput functional characterization of influenza viruses

机译：模块化糖球测定可用于流感病毒的高通量功能表征

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Background The ongoing global efforts to control influenza epidemics and pandemics require high-throughput technologies to detect, quantify, and functionally characterize viral isolates. The 2009 influenza pandemic as well as the recent in-vitro selection of highly transmissible H5N1 variants have only increased existing concerns about emerging influenza strains with significantly enhanced human-to-human transmissibility. High-affinity binding of the virus hemagglutinin to human receptor glycans is a highly sensitive and stringent indicator of host adaptation and virus transmissibility. The surveillance of receptor-binding characteristics can therefore provide a strong additional indicator for the relative hazard imposed by circulating and newly emerging influenza strains. Results Streptavidin -coated microspheres were coated with selected biotinylated glycans to mimic either human or avian influenza host-cell receptors. Such glycospheres were used to selectively capture influenza virus of diverse subtypes from a variety of samples. Bound virus was then detected by fluorescently labelled antibodies and analyzed by quantitative flow cytometry. Recombinant hemagglutinin, inactivated virus, and influenza virions were captured and analyzed with regards to receptor specificity over a wide range of analyte concentration. High-throughput analyses of influenza virus produced dose–response curves that allow for functional assessment of relative receptor affinity and thus transmissibility. Conclusions Modular glycosphere assays for high-throughput functional characterization of influenza viruses introduce an important tool to augment the surveillance of clinical and veterinarian influenza isolates with regards to receptor specificity, host adaptation, and virus transmissibility.
机译：背景技术为了控制流感流行和大流行，全球正在进行的努力需要高通量技术来检测，量化和功能鉴定病毒分离株。 2009年的流感大流行以及最近在体外选择的高传播性H5N1变体，仅增加了人们对新兴流感病毒株的担忧，这些病毒株具有明显的人对人传播能力。病毒血凝素与人受体聚糖的高亲和力结合是宿主适应性和病毒传播性的高度敏感和严格的指标。因此，受体结合特征的监测可以为循环和新出现的流感病毒株带来的相对危害提供强有力的附加指标。结果用抗生蛋白链菌素包被的微球用选定的生物素化聚糖包被，以模拟人或禽流感宿主细胞受体。此类糖球用于从多种样品中选择性捕获多种亚型的流感病毒。然后通过荧光标记的抗体检测结合的病毒，并通过定量流式细胞术进行分析。捕获重组血凝素，灭活病毒和流感病毒粒子，并在广泛的分析物浓度范围内就受体特异性进行分析。流感病毒的高通量分析产生了剂量反应曲线，可以对相对受体亲和力和因此的可传递性进行功能评估。结论用于流感病毒高通量功能表征的模块化糖球测定法在受体特异性，宿主适应性和病毒传播性方面引入了重要的工具，可增强对临床和兽医分离株的监测。

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